Vigna Δ¹-pyrroline-5-carboxylate synthetase (P5CS) cDNA was transferred to chickpea (Cicer arietinum L.) cultivar Annigeri via Agrobacterium tumefaciens mediated transformation. Following selection on hygromycin and regeneration, 60 hygromycin-resistant plants were recovered. Southern blot analysis of five fertile independent lines of T0 and T1 generation revealed single and multiple insertions of the transgene. RT-PCR and Western blot analysis of T0 and T1 progeny demonstrated that the P5CS gene is expressed and produced functional protein in chickpea. T1 transgenic lines accumulated higher amount of proline under 250 mM NaCl compared to untransformed controls. Higher accumulation of Na⁺ was noticed in the older leaves but negligible accumulation in seeds of T1 transgenic lines as compared to the controls. Chlorophyll stability and electrolyte leakage indicated that proline overproduction helps in alleviating salt stress in transgenic chickpea plants. The T1 transgenics lines were grown to maturity and set normal viable seeds under continuous salinity stress (250 mM) without any reduction in plant yield in terms of seed mass.

The transformation of potato (Solanum tuberosum L. cv. Désirée) was extended by the Agrobacterium-mediated biolistic method. Using this approach transgenic shoots could be obtained at a similar frequency to that achieved through conventional biolistics. Leaves from shoot cultures were bombarded with gold particles coated in Agrobacterium tumefaciens cells harboring a binary plasmid encoding three genes of interest in the T-DNA. Nine shoots were obtained from 20 shots, with selection of transgenic shoots on a series of media containing progressively increasing concentrations of hygromycin from 5 to 20 mg dm^-3.

Hexokinase (HXK, EC 2.7.1.1) plays an important role in the metabolism and glucose signalling. To examine the characteristics of HXK gene family in rice, the subcellular localizations of ten hexokinases (OsHXK1 - OsHXK10) were determined using OsHXK::GFP fusion proteins in tobacco mesophyll protoplasts. As was previously demonstrated, OsHXK4 was detected in the chloroplast stroma, OsHXK5 and OsHXK6 in the mitochondria, and OsHXK7 and OsHXK10 in the cytoplasm. In the present study, OsHXKs were clearly divided into three types (A, B, C) based on their N-terminal sequences. The new type-C HXKs in plants, OsHXK1, OsHXK7 and OsHXK8, which lack the plastidic transit peptide and the membrane anchor domain, were detected not only in the cytoplasm but also in the nucleus. The type-B HXKs, OsHXK2, OsHXK3, OsHXK9 and OsHXK10, which contained a membrane anchor domain, were distinctly localized in the mitochondria. These results suggest that OsHXKs localized in different cell compartments may be involved in the glucose signalling-related gene expression during growth and development of rice.

Mature seed-derived embryogenic calli of indica rice (Oryza sativa L. cv. PAU201) were induced on semisolid Murashige and Skoog medium supplemented with 2.5 mg dm^-3 2,4-dichlorophenoxyacetic acid + 0.5 mg dm^-3 kinetin + 560 mg dm^-3 proline + 30 g dm^-3 sucrose + 8 g dm^-3 agar. Using OsglyII gene, out of 3180 calli bombarded, 32 plants were regenerated on medium containing hygromycin (30 mg dm^-3). Histochemical GUS assay of the hygromycin selected calli revealed GUS expression in 50 % calli. Among the regenerants, 46.87 % were GUS positive. PCR analysis confirmed the presence of the transgene of 1 kb in 60 % of independent plants. Further, these plants have been grown to maturity in glasshouse. In vitro screening for salt tolerance showed increase in fresh mass of OsglyII putative transgenic calli (185.4 mg) as compared to control calli (84.2 mg) on 90 mM NaCl after 15 d. When exposed to 150 mM NaCl, OsglyII putative transgenic plantlets showed normal growth while the non-transgenic control plantlets turned yellow and finally did not survive.

The paper reports the in vitro cultivation of two commercial lines and 23 wild populations (with 10, 20 and 30 chromosomes) of Brachypodium distachyon. Callus induction was assayed on Murashige and Skoog medium containing 1 mg dm^-3 2,4-dichlorophenoxyacetic acid (2,4-D) with 30 g dm^-3 of sucrose (MSs) or maltose (MSm). No significant differences were seen between the two media with respect to callus induction. Calli were transferred to MSm medium without 2,4-D but containing 0.1 mg dm^-3 of 6-benzylaminopurine for plant regeneration. The plant regeneration response was very variable depending on the original induction medium, although no overall preference for one or the other medium was seen. The three main culture stages (callus induction, plant regeneration, and green plantlets formation) are probably differently controlled in the plants with different chromosome numbers. This supports the idea that the three cytotypes of Brachypodium cultured actually belong to different species.

Short-time direct and indirect effects of 25 μM Cd on the growth of dicotyledon (Phaseolus coccineus) and monocotyledon (Allium cepa) plants were investigated in the presence of inhibitors of ethylene synthesis, NADPH oxidase, and the octadecanoid pathway. Only 5 min-long action of Cd was enough for inhibition of growth in bean roots, but its recovery time was extended to several days. After 7 h treatment, Cd was significantly accumulated in bean roots, but maximum H₂O₂ accumulation was seen after 1 h. Cd-induced H₂O₂ accumulation decreased especially after addition of ethylene inhibitor silver thiosulphate (STS). Low Cd accumulation and high growth inhibition were observed also in bean leaves and in A. cepa roots. The inhibitors of the octadecanoid pathway greatly weakened the inhibitory effect of Cd in P. coccineus roots, while no significant effect was observed in A. cepa. NADPH oxidase and ethylene blockade reversed (in the case of bean plants and indirectly treated A. cepa plants) or significantly diminished Cd action. Cd-induced growth inhibition of P. coccineus leaves was also alleviated by most inhibitors of the jasmonate pathway and by STS. These results indicate that Cd may have indirect and direct effects on growth processes.

The study was conducted to investigate the physiological effects of exogenous NO on potherb mustard (Brassica juncea Coss.) seedlings under salt stress. The plants were grown in Hogland nutrient solution for 15 d and treated with 150 mM NaCl, NO donor sodium nitropruside (SNP) and NO scavenger methylene blue (MB-1) for 4 d. The NaCl stress increased superoxide dismutase, peroxidase and ascorbate peroxidase activities and malondialdehyde (MDA) and free proline contents, and decreased soluble protein content. However, the application of exogenous NO limited the production of MDA and free proline, while markedly promoted SOD, POD and APX activity.

The objective of this study was to examine the role of NADPH oxidase on superoxide radical production under waterlogging in mung bean (Vigna radiata) cvs. T 44 (tolerant) and Pusa Baisakhi (PB) (susceptible), and wild species Vigna luteola. Two days of waterlogging caused decline in superoxide radical (O₂ ^.-) contents in all the genotypes, however, further waterlogging up to 8 d caused significant increase in O₂ ^.- contents. In control and revived plants O₂ ^.- contents were higher in PB, while under waterlogging stress T 44 and V. luteola showed greater increases in the O₂ ^.- contents. During waterlogging the increase in O₂ ^.- content was found to be due to the diphenylene iodonium chloridesensitive NADPH oxidase (NOX). This was further confirmed by the waterlogging induced increase in NOX activity, which was higher in tolerant genotypes T 44 and V. luteola compared with PB. Gene expression studies showed enhanced expression of NOX in the roots of waterlogged V. luteola and T 44, while little expression was observed in control or treated plants of PB. PCR band products were cloned and sequenced, and partial cDNAs of NOX was obtained. Results suggest that increase in O₂ ^.- content during waterlogging could be due to the induction of membrane linked NOX.

Biotechnological improvement of monocots is often hampered by the lack of efficient regeneration systems, requisite wound responses and low cell competence. Despite these limitations, the biolistic and Agrobacterium methods have been successfully used to produce several transgenic monocots by adjusting the parameters that govern efficient delivery and integration of transgene(s) into plant genome. It is now possible to transform even difficult monocots using tailor-made gene constructs and promoters, suitable A. tumefaciens strains and a proper understanding of the entire process. This success has been reviewed in the present article and a special emphasis was laid on the measures that were taken in overcoming the difficulties that arise due to the differential responses of monocots and dicots. This information is necessary for biotechnological improvement of still newer monocotyledonous plants that have been hitherto difficult to transform.

The effect of aluminum and chromium on two barley genotypes differing in Al tolerance was studied in a hydroponic experiment. Al stress decreased plant growth, biomass production, chlorophyll content and photosynthetic efficiency determined as variable to maximum chlorophyll fluorescence ratio (F_v/F_m), net photosynthetic rate (P_N), intercellular CO₂ concentration (c_i), stomatal conductance (g_s) and transpiration rate (E) less in an Al-tolerant genotype Gebeina than in an Al-sensitive genotype Shang 70-119. Cr stress also caused marked reduction in growth and photosynthetic traits in barley plants. Higher reduction was observed at pH 4.0 as compared to pH 6.5. Combined stress of Cr and Al, caused further reduction in growth and photosynthetic parameters.

The elucidation of molecular mechanisms underlying the leaf development can be facilitated by the detailed anatomical study of leaf development mutants. We present an analysis of leaf anatomy and morphogenesis during early developmental stages in has mutant of Arabidopsis thaliana. The recessive has mutation affects a number of aspects in plant development, including the shape and size of both cotyledons and leaves. The earliest developmental observations suggest almost synchronous growth of the first two leaf primordia of has mutant. No significant disruption of the cell division pattern in the internal tissue is observed at the earliest stages of development, with the major anatomical difference compared to wild type primordia being the untimely maturation of mesophyll tissue cells in has mutant. At the stage of leaf blade formation, structure disruption becomes clearly evident, by irregular arrangement of the cell layers and the lack of polarity in juvenile has leaves. One distinguishing feature of the mutant leaf anatomy is the absence of mesophyll tissue differentiation. Altered has mutant leaf morphology could be at least partially accounted for by the ectopic STM activity that was found at the base of leaf primordia during early stages of leaf development in has plants.

The structural features of flavonoids which are involved in the modulation of auxin distribution in Arabidopsis thaliana were evaluated. An auxin-inducible promoter IAA2 fused to a reporter gene (GUS) was used to monitor the tissue responsiveness to auxins. The following aspects were investigated: 1) the influence of flavonoids (quercetin, naringenin, kaempferol, myricetin and isorhamnetin) on the distribution of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) in roots and leaves, 2) differences in flavonoid uptake into roots and shoots depending on flavonoid concentration in the medium, and 3) influence of structurally different flavonoids on the gravitropic response and growth of roots. The same flavonoids differently affected IAA and IBA distribution in leaves and roots. There were several structural requirements for the flavonoids which resulted in the changes of auxin response/distribution. Great differences between the ability of shoots and roots to take up quercetin were showed. Also, flavonoids influenced gravitropism and root growth of Arabidopsis seedlings in a structure-dependent manner.

Pea (Pisum sativum L., cv. Scinado) seedlings were exposed to low doses of ultraviolet-B (UV-B; 4.4 and 13.3 kJ m^-2 d^-1) or UV-C (0.1 and 0.3 kJ m^-2 d^-1) radiation for 14 d. Aminocyclopropane carboxylic acid (ACC), indoleacetic acid (IAA) and abscisic acid (ABA) contents were quantified by gas chromatography coupled to mass spectrometry (GC-MS). The accumulation of ACC upon irradiation was dose-dependent. ABA content was reduced and IAA content increased upon UV-C treatment whereas the UV-B doses used did not cause significant changes in ABA and IAA contents.

A rapid and highly-effective method for micropropagation from nodal segment and shoot tip explants was established for Coleus blumei Benth. Nodal segments and shoot tips were inoculated on MS medium containing 0.7 % agar, 3 % commercial sugar, and different combinations of 6-benzyladenine (BA) with indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) or α-naphthaleneacetic acid (NAA). Hundred percent shoot induction from both explants was achieved on the medium containing BA (2 mg dm^-3) and NAA (1 mg dm^-3). Shoot tips were proved to be the better explant in comparison to nodal segments in having high rate of shoot induction and more number of shoots. The same media conditions were found suitable for shoot multiplication. Multiplied shoots rooted best on MS medium supplemented with IBA (2 mg dm^-3). Micropropagated plants were successfully established in soil after hardening, with 100 % survival rate.

A reproducible protocol has been developed for high frequency plant regeneration from immature embryos of Argyrolobium roseum Jaub & Spach, an important medicinal legume. Green nodular calli were initiated from immature embryos excised from 10-d-old pods in 70 % of cultures within 3 weeks when grown on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm^-3 benzylaminopurine (BAP) + 0.25 mg dm^-3 indole-3-acetic acid (IAA). Subsequent transfer of 5 mm² callus pieces to MS medium supplemented with BAP (0.5 mg dm^-3) alone or in combination with IAA (0.25 mg dm^-3) facilitated regeneration of multiple shoots. Organogenic calli bearing multiple shoots when transferred to MS medium supplemented with BAP (0.5 mg dm^-3) + IAA (0.25 mg dm^-3) supported rapid shoot elongation. Shoot propagules subcultured to Gamborg's medium (B5) with 0.5 mg dm^-3 indole-3-butyric acid (IBA) rooted with 80 % frequency and developed into phenotypically normal plants. Plantlets were successfully acclimatized in a sterile mixture of sand and garden soil (1:1) under greenhouse and thereafter transferred to field beds.

Genetic engineering of rice (Oryza sativa L. cv. Pusa basmati 1) using synthetic Cry1Ac gene has been achieved by "particle bombardment". Scutellar tissues excised after 5 - 6 d from mature seeds cultured on induction medium were bombarded using gold particles coated with a mixture of Cry1Ac and marker genes on medium with osmoticum. Bombarded tissues were subjected to 30 mg dm^-3 hygromycin selection for two cycles. The selected calli after GUS assay were transferred to shoot regeneration medium. Regenerated shoots were rooted and plantlets (T₀) were grown to full maturity. Polymerase chain reaction (PCR) analysis of T₀ plants using Cry1Ac specific primers revealed the presence of Cry1Ac gene in 65 % plants. Phenotypic assay, β-glucuronidase assay and PCR during T₁ generation revealed the inheritance of the Cry1Ac and marker genes along with the native plant genes.

In red clover (Trifolium pratense L.) four unique cDNAs encoding phenylalanine ammonia lyase (PAL, EC 4.3.1.5) were identified (PAL1-4). PAL2-4 encode nearly identical proteins (> 97 %) that are only 89 % identical to that encoded by PAL1. Under normal growing conditions, in young leaves and flowers PAL1 mRNA accumulates to higher levels than that of PAL2-4 whereas in mature leaves and stems, mRNA levels are similar for PAL1 and PAL2-4. Treatment of red clover seedlings with yeast elicitor preparation results in an approximately six-fold induction of PAL2-4 transcripts within 1 h of treatment but only a modest induction of PAL1 transcripts. These results suggest that while both classes of enzymes play a role in biosynthesis of phenylpropanoid compounds under normal growing conditions, PAL2-4 enzymes are also involved in pathogen defense responses.

Multiple shoot buds could be induced directly from internode explants of Celastrus paniculatus inoculated on Murashige and Skoog's (MS) medium containing different growth regulators. The best response was obtained when 4.44 μM 6-benzylaminopurine (BAP) was incorporated in the medium. Incorporation of indole-3-acetic acid (IAA) and α-naphthalene acetic acid (NAA) did not improve response, rather promoted callusing. Adventitious shoot buds could be multiplied and elongated on MS medium containing 2.22 μM BAP. Rooting of shoots (80 %) was obtained when their bases were dipped in pre-autoclaved indole-3-butyric acid (IBA) solution (2.45 mM) for 10 min followed by their implantation on medium containing 1/4 MS salts, 1.0 % sucrose and 0.6 % agar. Out of 500 plantlets subjected to hardening, 410 were successfully hardened under greenhouse conditions. Twenty plants were established in field while remaining of them were transferred to nursery conditions without any mortality.

The emissions of volatile organic compounds, VOC, from plants have strong relevance for plant physiology, plant ecology, and atmospheric chemistry. We report here on the current knowledge of the many internal (genetic and biochemical) and external (abiotic - temperature, light, water availability, wind, ozone, and biotic - animal, plant and microorganisms interactions) factors that control emission rates of different VOC by altering their synthesis, vapour pressure or diffusion to the atmosphere. The complex net of these factors, their interactions and the different responses of the different VOC produces the large qualitative and quantitative, spatial and temporal variability of emissions and the frequent deviations from current standard emission models. The need for a co-operative multidisciplinary multiscale research to disentangle this complex and important issue of plant VOC emissions is reminded.

The aim of this study was to investigate the protein alterations in the transgenic kiwifruit (Actinidia deliciosa) overexpressing a transcription factor gene, OSH1. Although transgenic plant with introduced OSH1 indicates suppression of gibberellin (GA)-20 oxidase activity, application of GA to transgenic kiwifruit could not completely recover plant morphology and protein profiles. Eleven proteins decreased in the transgenic kiwifruit detected by two-dimensional polyacrylamide gel electrophoresis showed homologies to kiwifruit hypothetical protein, osmotin I and photosynthesis related protein. These results suggest that introduction of an OSH1 into kiwifruit caused wide-range alterations at protein level and alterations of protein accumulation should be considered to evaluate the substantial equivalence of plants transformed by a transcription factor.

Hairy root cultures of Hypericum perforatum were obtained following inoculation of aseptically germinated seedlings with A. rhizogenes strain A4M70GUS. Effect of sucrose on the growth and biomass production of hairy root cultures was investigated. Hairy root cultures spontaneously regenerated shoots buds from which a number of shoot culture clones was established. Transformed shoot cultures exhibited good shoot multiplication, elongation and rooting on a hormone-free woody plant medium. Plants regenerated from hairy roots were similar in appearance to the normal, nontransformed plants.

The allelopathic potential of four tree species on soil microbial populations and some herbaceous plants (two understory species and one general biotest species) was investigated. Effects of three nonindigenous tree species, Eucalyptus globulus Labill, Pinus radiata D.Don and Acacia melanoxylon R.Br., on microorganisms participating in the cycle of nitrogen were evaluated, comparing them with those produced by the autochthonous Quercus robur L. Influence of the trees on Lactuca sativa L., Dactylis glomerata L. and Trifolium repens L. was also checked in bioassays. Cell numbers of Nitrosomonas sp. were negatively affected by Acacia and Eucalyptus stands, mainly during spring, when flowers are especially abundant on the ground. Proteolytic microorganisms were also negatively affected by Eucalyptus and Pinus stands, whilst Quercus stand did not show any toxicity. Soil bioassays showed clear inhibitory effects on germination and growth of understory plants, particularly soils from Eucalyptus and Acacia stands. The greatest effects had the soil from Acacia stand, which was phytotoxic during the whole period of germination and growth of understory plants. Allelopathic phenomena could be, at least partially, responsible of the low species diversity in the understory of the nonindigenous tree stands.

Three clones of tobacco transformed with the T-DNA of Agrobacterium rhizogenes Ri plasmid A4b cultivated in vitro on a solid agar medium neither showed pronounced morphological diversity nor significantly differed in chlorophyll (Chl) contents from control plants. However, the transformation caused a 27 to 83 % decay in leaf oxygen evolution and in both ribulose-1,5-bisphosphate carboxylase (RuBPC) and phosphoenolpyruvate carboxylase (PEPC) activities. Therefore, the transformation brought about a reduction of active PEPC as well as activable RuBPC amounts in plant tissues. Individual clones did not mutually differ. In tobacco transformed with A. rhizogenes strain TR101 and grown in soil only the mean leaf area tended to reduce. Chl contents, Chl a/b ratio, oxygen evolution, and activities of both RuBPC and PEPC were insignificantly affected by the transformation.

Susceptibility of Mexican cacti to Agrobacterium rhizogenes was evaluated in 65 species of 22 genera. Stem discs taken from in vitro cultured plants were inoculated with Agrobacterium rhizogenes A4 agropine-type strain that contains the wild RiA4 plasmid and the binary vector pESC4 with the nptII and gus genes. Hairy roots were produced directly from wounds, or starting from calli generated on the wounded surface, in 34 of the evaluated species. The frequency of hairy roots formation, the number of roots per explant and its growth rates were variable among the tested species. In the 31 remaining species the production of transformed roots was not observed under the conditions used in these experiments. Histochemical detection of β-glucuronidase (GUS) activity demonstrated the expression of this foreign gene in the hairy roots. PCR analyses demonstrated the presence of the rolB and nptII genes in the DNA of the transformed roots. The patterns of alkaloid-like compounds obtained by thin layer chromatography in some of the tested species were qualitatively similar between the transformed and non-transformed roots.

Cytokinin (CK) content and activities of several antioxidant enzymes were examined during plant ontogeny with the aim to elucidate their role in delayed senescence of transgenic Pssu-ipt tobacco. Control Nicotiana tabacum L. (cv. Petit Havana SR1) and transgenic tobacco with the ipt gene under the control of the promoter of small subunit of Rubisco (Pssu-ipt) were both grown either as grafts on control rootstocks or as rooted plants. Both control plant types showed a decline in total content of CKs with proceeding plant senescence. Contrary to this both transgenic plant types exhibited at least ten times higher content of CKs than controls and a significant increase of CK contents throughout the ontogeny with maximal values in the later stages of plant development. Significantly higher portion of O-glucosides was found in both transgenic plant types compared to control ones. In transgenic plants, zeatin and zeatin riboside were predominant type of CKs. Generally, Pssu-ipt tobacco exhibited elevated activities of antioxidant enzymes compared to control tobacco particularly in the later stages of plant development. While in control tobacco activity of glutathione reductase (GR) and superoxide dismutase (SOD) showed increasing activity up to the onset of flowering and then gradually decreased, in both transgenic types GR increased and SOD activity showed only small change throughout the plant ontogeny. Ascorbate peroxidase (APOD) was stimulated in both transgenic types. The manifold enhancement of syringaldazine and guaiacol peroxidase activities was observed in transgenic grafts throughout plant ontogeny in contrast to control and transgenic rooted plants, where the increase was found only in the late stages. Electron microscopic examination showed higher number of crystallic cores in peroxisomes and abnormal interactions among organelles in transgenic tobacco in comparison with control plant. The overproduction of cytokinins resulted in the stimulation of activities of AOE throughout the plant ontogeny of transgenic Pssu-ipt tobacco.

The photosynthetic performance and related leaf traits of Incarvillea delavayi Bur. et Franch were studied at different water regimes to assess its capacity for photosynthetic acclimation to water stress. The initial response of I. delavayi to water stress was the closure of stomata, which resulted in down-regulation of photosynthesis. The stomatal limitation (S_L) represented the main component to photosynthetic limitations but non-stomatal limitation (NS_L) increased quickly with the increasing water stress, and had similar magnitude to SL under severe water stress (soil moisture 25-30 % of field capacity). Chlorophyll (Chl) a fluorescence parameters characterizing photosystem (PS) 2 photochemical efficiency (Φ_PS2), electron transport rate (J) and photochemical quenching (qP) decreased with the increasing water stress, indicating impaired photosynthetic apparatus. However, the water-stressed plants had a increased mesophyll CO₂ diffusional conductance, Chl a/b ratio, leaf nitrogen partitioning in RuBPCO and bioenergetics in later grown parts, indicating that I. delavay had a substantial physiological plasticity and showed a good tolerance to water stress.

The effect of 5-azacytidine (5-azaC) on the alleviation of damaging effects of NaCl treatment was studied in two wheat (Triticum aestivum L.) cultivars differing in salt tolerance (salt-tolerant Dekang-961 and sensitive Lumai-15). The plants were pre-treated or not with 50 μM 5-azaC and then subjected to salt stress induced by 100 or 150 mM NaCl. Salinity caused reduction in biomass accumulation and increase in malondialdehyde content in root tissues in both cultivars, but less in pre-treated seedlings. The activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in the roots of both cultivars increased during salt stress, but the rate of increase was higher in Dekang-961. Plants treated with 5-azaC had higher root SOD, CAT and POD activities under salt stress than untreated plants. Content of 5-methylcytosine (5mC) decreased in both cultivars under salt stress, and the level of demethylation was higher in Dekang-961 than that in Lumai-15. Moreover, the degree of methylation was lower in both cultivars under salt stress after 5-azaC application compared to only salt-treated groups. These findings suggested that 5-azaC could protect plants from salt stress.

In this communication, we report the binding of abscisic acid responsive elements (ABREs) of rice Osem, namely motif A and motif B, with a cognate trans-acting factor present in the nuclear extract of tobacco leaf. The binding is specific as both the complexes were disrupted with an excess of homologous non-radioactive DNA like motif A or motif B themselves or with cis-elements of rice Rab16A, motif I (ABRE) and motif IIa (non-ACGT ABRE-like sequences). Four tandem repeats of ABRE from wheat Em (4X ABRE) or two tandem repeats of Em ABRE, plus two copies of coupling element (CE1) from barley HVA22 (2X ABRC), also showed specific complexes, that were competed out by an excess of homologous competitors like motif I, motif IIa, motif A, motif B, 4X ABRE and 2X ABRC, but not by the unrelated 4X DRE sequence. Elution of the protein from all the complexes showed a single 26 kDa polypeptide band. Introgression of two of the above synthetic promoters 4X ABRE and 2X ABRC, each fused with minimal promoter of cauliflower mosaic virus 35S (CaMV 35S), could induce the expression of the reporter gene β-glucuronidase (gus) in transgenic tobacco in response to high NaCl concentration, dehydration or abscisic acid, but not at the constitutive level, proving that they can be used as efficient stress-inducible promoters. Our work shows both in vivo and in vitro activity of the promoters from monocot genes in the model dicot plant tobacco.

Effects of the infection with tobacco mosaic virus (TMV) and potato virus Y (PVY) on chloroplasts from susceptible tobacco plants were determined. Changes in ribonucleases (RNases), phosphomonoesterase (PME), phosphodiesterase (PDE), glucose-6-phosphate dehydrogenase (G6P DH), 6-phosphogluconate dehydrogenase (6PG DH), glucokinase (GK), and fructokinase (FK) activities in thylakoid/envelope and stroma fractions were studied. Slight increase in the activities of PME, PDE, G6P DH and 6PG DH of thylakoid/envelope fraction as well as of RNases, PME, PDE, G6P DH, 6PG DH, GK and FK of stroma fraction was found in chloroplasts isolated from leaf tissues infected with PVY. Infection with TMV produced higher increase in enzymes activities in chloroplasts; especially, PME, G6P DH and 6PG DH in fraction of thylakoid/envelope, and RNases, PME, PDE, G6P DH, 6PG DH, and GK in stroma fraction.